Targeted Single-Cell RNA-seq Identifies Minority Cell Types of Kidney Distal Nephron
Targeted Single-Cell RNA-seq Identifies Minority Cell Types of Kidney Distal Nephron
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DOI：
10.1681/asn.2020101407
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发表时间：
2021-04-01
影响因子：
13.6
通讯作者：
Knepper, Mark A.
中科院分区：
文献类型：
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作者：
Chen, Lihe;Chou, Chun-Lin;Knepper, Mark A.
背景近端肾小管细胞在数量上占主导地位，尽管远端肾单位的细胞类型在水和电解质平衡中的作用不成比例。方法基于FACS的富集法与单细胞RNA-SEQ相结合，分析了9099个小鼠肾粗升支(CTAL)/远端曲管(DCT)区域细胞的转录本。结果非监督聚类发现SLc12a3(+)/Pvalb(+)和slc12a3(+)/Pvalb(-)细胞，分别被鉴定为DCT1和DCT2细胞。DCT1细胞呈异质性，SLC8a1、Calb1和CkB的表达呈正交性变化。另外一个DCT1亚群显示与细胞周期/细胞增殖相关的mRNAs显著丰富(例如，Mki67、STMN1和Top2a)，这符合DCT细胞的已知可塑性。没有发现DCT2特异的转录本。DCT2细胞与DCT1细胞相比，上皮钠通道β和伽马亚基的表达以及与钙转运相关的转录产物(TRPV5、Calb1、S100g和SLc8a1)的表达要强得多。此外，scRNA-seq鉴定了三种不同的CTAL(SLc12a1(+))细胞亚型。其中一个细胞表达NOS1和AVPR1A，与致密斑细胞一致。另外两个CTAL簇分别是Cldn10和Ptger3，以及Cldn16和Foxq1。在Cldn101和Cldn16(+)CTAL细胞中表达IRX1和IRX2，在Cldn16(+)CTAL细胞中表达IRX3。结论单细胞转录切割技术揭示了小鼠远端肾单位细胞间的差异。为单元格数据提供基于Web的数据资源。
Background Proximal tubule cells dominate the kidney parenchyma numerically, although less abundant cell types of the distal nephron have disproportionate roles in water and electrolyte balance.Methods Coupling of a FACS-based enrichment protocol with single-cell RNA-seq profiled the transcriptomes of 9099 cells from the thick ascending limb (CTAL)/distal convoluted tubule (DCT) region of the mouse nephron.Results Unsupervised clustering revealed Slc12a3(+)/ Pvalb(+) and Slc12a3(+)/Pvalb(-) cells, identified as DCT1 and DCT2 cells, respectively. DCT1 cells appear to be heterogeneous, with orthogonally variable expression of Slc8a1, Calb1, and Ckb. An additional DCT1 subcluster showed marked enrichment of cell cycle-/ cell proliferation-associated mRNAs (e.g., Mki67, Stmn1, andTop2a), which fitwith the known plasticity of DCT cells. No DCT2-specific transcripts were found. DCT2 cells contrast with DCT1 cells by expression of epithelial sodium channel beta- and gamma-subunits and much stronger expression of transcripts associated with calcium transport (Trpv5, Calb1, S100g, and Slc8a1). Additionally, scRNA-seq identified three distinct CTAL (Slc12a1(+)) cell subtypes. One of these expressed Nos1 and Avpr1a, consistent with macula densa cells. The other two CTAL clusters were distinguished by Cldn10 and Ptger3 in one and Cldn16 and Foxq1 in the other. These two CTAL cell types were also distinguished by expression of alternative Iroquois homeobox transcription factors, with Irx1 and Irx2 in the Cldn101 CTAL cells and Irx3 in the Cldn16(+) CTAL cells.Conclusions Single-cell transcriptomics revealed unexpected diversity among the cells of the distal nephron in mouse. Web-based data resources are provided for the single-cell data.